Small-volume cryogenic storage container

ABSTRACT

A cryovial device is disclosed including a vial configured to hold a liquid sample and an inlet/outlet tube coupled to the vial. The inlet/outlet tube is constructed of a weldable polymer and has a filled configuration, a closed configuration, and a drained configuration.

REFERENCE TO RELATED APPLICATIONS

This application claims an invention disclosed in U.S. ProvisionalApplication No. 63/218,550, filed Jul. 6, 2021, entitled “Small-VolumeCryogenic Storage Container”. Benefit under 35 USC § 119(e) of theUnited States provisional application is claimed, and the aforementionedapplication is incorporated herein by reference.

FIELD OF THE DISCLOSURE

The present disclosure relates to cryopreservation. More particularly,the present disclosure relates to a cryovial device and to a method forusing the same.

BACKGROUND OF THE DISCLOSURE

Cryopreservation is the process of cooling and storing biologicalmaterial (e.g., cells, tissues, organs) at very low temperatures tomaintain their viability for future use. The biological material'spost-thaw function should be sufficiently representative of thebiological material's pre-freeze function.

Cryovials are commonly used for cryopreservation. Such cryovials shouldbe capable of withstanding cryogenic temperatures while also avoidingcontamination or leakage of the biological material. Such cryovialsshould also be efficient and compatible for use in different laboratoryand clinical settings.

SUMMARY

A cryovial device is disclosed including a vial configured to hold aliquid sample and an inlet/outlet tube coupled to the vial. Theinlet/outlet tube is constructed of a weldable polymer and has a filledconfiguration, a closed configuration, and a drained configuration.

According to an exemplary embodiment of the present disclosure, acryovial device is disclosed including a vial configured to hold aliquid sample, an inlet/outlet tube coupled to the vial and constructedof a weldable polymer, the inlet/outlet tube having a filledconfiguration in which the inlet/outlet tube is coupled to a source ofthe liquid sample, and a drained configuration in which the inlet/outlettube is coupled to a receiving tube, and a vent tube coupled to thevial.

According to another exemplary embodiment of the present disclosure, amethod of using a cryovial device is disclosed including a vial and aninlet/outlet tube. The method includes the steps of filling the vialwith a liquid sample via the inlet/outlet tube, closing the inlet/outlettube after the filling step, cryopreserving the sample in the vial afterthe closing step, opening the inlet/outlet tube after the cryopreservingstep, coupling the inlet/outlet tube to a receiving tube, and drainingthe sample from the vial into the receiving tube via the inlet/outlettube.

BRIEF DESCRIPTION OF THE DRAWINGS

The above-mentioned and other features and advantages of thisdisclosure, and the manner of attaining them, will become more apparentand will be better understood by reference to the following descriptionof embodiments of the invention taken in conjunction with theaccompanying drawings, wherein:

FIG. 1 is a perspective view of an exemplary cryovial device of thepresent disclosure;

FIG. 2 is a front elevational view of the cryovial device of FIG. 1 ;

FIG. 3 is a side elevational view of the cryovial device of FIG. 1 ;

FIG. 4 is a perspective view of an optionally used seal elementunassembled with the cryovial device of FIG. 1 .

FIG. 5 is a perspective view of the seal element of FIG. 4 assembled tothe cryovial device of FIG. 1 , with a portion of the cryovial device ofFIG. 1 removed to view the seal element of FIG. 4 .

FIG. 6 is a perspective view of the cryovial device of FIG. 1additionally including the seal element of FIG. 4 .

FIG. 7 is an elevational view of a storage container for holding one ormore of the cryovial devices of FIG. 1 ; and

FIG. 8 shows a method of using the cryovial device of FIG. 1 , themethod including a filling step (a), a closing step (b), a severing step(c), an unraveling step (d), an opening step (e), a coupling step (f),and a draining step (g).

Corresponding reference characters indicate corresponding partsthroughout the several views. The exemplifications set out hereinillustrate exemplary embodiments of the invention and suchexemplifications are not to be construed as limiting the scope of theinvention in any manner.

DETAILED DESCRIPTION Cryovial Device

A cryovial device 100 is shown in FIGS. 1-3 . The cryovial device 100 isconfigured to receive a liquid sample, contain the sample duringcryostorage, and deliver the thawed sample. The sample may include abiological fluid, such as a suspension of blood cells (e.g.,hematopoietic stem and progenitor cells (HPCs) derived from prematurecord blood (PCB)). The sample may also include electrolytes and/orcryoprotectants (e.g., glycerol, propylene glycol, ethylene glycol,dimethyl sulfoxide (DMSO)). The cryovial device 100 may be considered asubstantially closed system with fluid-tight materials and joints thatare capable of withstanding cryogenic temperatures (e.g., about −196°C.).

The illustrative cryovial device 100 of FIGS. 1-3 includes a vial 200, afirst, inlet/outlet tube 300, a second, vent tube 400, a tube clip 500,and a spool 600. Each element of the cryovial device 100 is describedfurther below.

The vial 200 of the illustrative cryovial device 100 is configured tocontain the sample. The illustrative vial 200 is configured to holdabout 2 mL to about 5 mL of the sample, although this volume may varyfrom about 1 mL to about 30 mL or more. The illustrative vial 200 iscylindrical in shape, although this shape may also vary. The vial 200has a closed lower end 202 and an upper end 204 with a first,inlet/outlet opening 205 and a second, vent opening 207. The first,inlet/outlet opening 205 is defined by a first fitting 206, which isconfigured to couple to the first, inlet/outlet tube 300. The second,vent opening 207 is defined by a second fitting 208, which is configuredto couple to the second, vent tube 400. The illustrative fittings 206,208 are barbed and configured to be friction-fit within their respectivetubes 300, 400, but it is also within the scope of the presentdisclosure for the fittings 206, 208 to be heat-sealed, molded, adhered,and/or otherwise coupled to their respective tubes 300, 400. The firstfitting 206 is illustratively taller than the second fitting 208,although this arrangement may vary. The vial 200 may be constructed of arigid material such as polystyrene, polypropylene, or another suitablematerial.

In some embodiments, a sealing element 210 can be used, as illustratedin FIGS. 4-6 , to facilitate securing the inlet/outlet tube 300 and thevent tube 400 to the respective fittings 206, 208, and to reduce orprevent fluid leakage between the tubes 300, 400 and their respectivefittings 206, 208. The sealing element 210 can have two holes 212 spacedin correspondence with the spacing between the inlet/outlet tube 300 andthe vent tube 400. The holes 212 can be of a size or diameter for afriction or interference fit around the inlet/outlet tube 300 and thevent tube 400, to squeeze or compress the tubes 300, 400 around theirrespective fittings 206, 208. The friction or interference fit can beaccomplished by fabricating the sealing element 210 from an elastomericmaterial that stretches and compresses around the tubes 300, 400, aheat-shrink material that shrinks to compress around the tubes 300, 400,or a non-elastomeric material such as plastic or metal.

Referring again to FIGS. 1-3 , the inlet/outlet tube 300 of theillustrative cryovial device 100 is configured to both receive theliquid sample and deliver the thawed sample through the inlet/outletopening 205 of the vial 200. In this way, the dual-purpose inlet/outlettube 300 and its corresponding, dual-purpose inlet/outlet opening 205may eliminate the need for distinct inlet and outlet openings in thevial 200. Initially, the inlet/outlet tube 300 may be provided with adesired fill port 302. The illustrative fill port 302 is a needle-free,female Luer fitting having a normally closed diaphragm valve that openswhen coupled to an industry-standard, male Luer fitting. However, thefill port 302 may vary based on the intended application. For example,the fill port 302 may include a needle septum configured to be piercedby a syringe needle. The first, inlet/outlet tube 300 may be longer thanthe second, vent tube 400, and this excess length may be wrapped aroundthe spool 600, as discussed further below. The inlet/outlet tube 300 maybe constructed of a flexible, pharmaceutical grade, weldable polymer.For example, the inlet/outlet tube 300 may be constructed of athermoplastic elastomer (TPE) tubing, such as Tygon® tubing availablefrom Saint-Gobain Performance Plastics.

The vent tube 400 of the illustrative cryovial device 100 is configuredto vent gas into and/or from the vial 200 through the second, ventopening 207 while remaining liquid-tight. For example, the vent tube 400may allow air to pass from the vial 200 during filling and into the vial200 during draining. The vent tube 400 includes a filter element 402along its length that is configured to filter the air entering the vial200 during draining and/or at other times. The illustrative filterelement 402 is positioned about midway along the length of the vent tube400 between a lower tube portion 404 and an upper tube portion 406,although the location of the filter element 402 may vary. The filterelement 402 may be a micro-filter, such as a 3 μm sterile micro-filter.The filter element 402 may be gas permeable but liquid impermeable toavoid leakage of the sample from the vial 200. The vent tube 400, likethe inlet/outlet tube 300, may be constructed of a flexible,pharmaceutical grade, thermoplastic elastomer (TPE) tubing, such asTygon® tubing available from Saint-Gobain Performance Plastics.

The tube clip 500 of the illustrative cryovial device 100 is configuredto support and stabilize the tubes 300, 400. The tube clip 500 may be a“3”-shaped component including a first recess 502 configured to hold thefirst, inlet/outlet tube 300, and a second recess 504 adjacent to thefirst recess 502 and configured to hold the second, vent tube 400. Thetube clip 500 may be sized to slide along the tubes 300, 400 and may bedetached from the tubes 300, 400, such as by pinching and removing thetubes 300, 400.

The spool 600 of the illustrative cryovial device 100 is configured tosupport and stabilize the first, inlet/outlet tube 300. The spool 600may be constructed of a first portion 602 and a second portion 604 thatare snap-fit together. The spool 600 may include a barrel 606 configuredto receive the first, inlet/outlet tube 300 in a coiled manner. Thespool 600 may also include a passageway 608 configured to freely receivethe second, vent tube 400.

Referring next to FIG. 7 , the illustrative cryovial device 100 may besized for receipt in a standard, tray-shaped, “egg carton” type storagecontainer 700 used to transfer and store cell samples for freezing andeventual thawing. For example, the vial 200 of the cryovial device 100may be sized for receipt in a separated area 702 of the storagecontainer 700 having a diameter of about 10 mm and a height of about 90mm. The tubes 300, 400 may project upward from the vial 200 and thestorage container 700, supported by the tube clip 500 and/or the spool600. As shown in FIG. 7 , several such cryovial devices 100,illustratively cryovial devices 100 a-100 d, carrying cell samples froma common source may be arranged in an array and housed in a commonstorage container 700.

Method of Use

An exemplary method of using the cryovial device 100 is demonstrated inFIG. 8 and described below. During some or all of the following steps,the vial 200 may be present in the above-described storage container 700(FIG. 7 ) with the tubes 300, 400 supported by the tube clip 500 and/orthe spool 600.

The method of FIG. 8 begins with a filling step (a) with the cryovialdevice 100 in a filled configuration. During this filling step (a), thesample is transferred from a source S, through the inlet/outlet tube300, and into the inlet/outlet opening 205 of the vial 200 (FIG. 1 ), asindicated by arrow A. The source S may be a syringe, a blood bag, oranother suitable container for the sample. In certain embodiments, thesource S may be present in an automated filling system, such as theCellSeal® AF-500™ filling system or the Signata CT-5™ filling system,both available from Sexton Biotechnologies. The source S may be coupled(e.g., Luer-locked) to the inlet/outlet tube 300 via the fill port 302,as shown in FIG. 8 . Alternatively, the fill port 302 may be removed,and the source S may be coupled (e.g., welded) to the inlet/outlet tube300 in a direct, closed manner. The sample may be introduced under theinfluence of gravity, positive pressure from the source S, and/or vacuumpressure through the vent tube 400. Air may escape from the vial 200 viathe vent tube 400 during this filling step (a).

The method of FIG. 8 continues with a closing step (b) with the cryovialdevice 100 in a closed configuration. During this closing step (b), theinlet/outlet tube 300 is heat-sealed or otherwise closed at seal 310 andthe vent tube 400 is heat-sealed or otherwise closed at seal 410 tocontain the sample in the cryovial device 100. The seal 310 may belocated between the first fitting 206 of the vial 200 (FIG. 1 ) and thefill port 302 of the inlet/outlet tube 300 and above the height of thevent tube 400 to avoid interfering with the vent tube 400. The seal 410may be located above the filter element 402 (FIG. 1 ) of the vent tube400. The closing step (b) may be performed using a medical-grade tubesealer that pinches and welds the inlet/outlet tube 300, such as theC'EAL-FLEX® TPE Ultra Sealer available from Saint-Gobain.

The method of FIG. 8 continues with a severing step (c) with thecryovial device 100 in a severed configuration. During this severingstep (c), the excess inlet/outlet tube 300 is sliced along cut line 312at or above the seal 310 and removed. This severing step (c) may beperformed substantially simultaneously with the above-described closingstep (b) in a closed environment. For example, both the closing step (b)and the severing step (c) may be performed using the above-describedtube sealer.

The method of FIG. 8 continues with an unraveling step (d) with thecryovial device 100 in an unraveled configuration. During thisunraveling step (d), the inlet/outlet tube 300 is unraveled from thespool 600 (FIG. 1 ), as indicated by arrow B. This unraveling step (d)gives the inlet/outlet tube 300 added length and clearance above thevent tube 400.

With the inlet/outlet tube 300 sealed, the sample in the cryovial device100 may be processed. For example, the sample may be cryogenicallyfrozen, stored/banked, and eventually thawed. It is also within thescope of the present disclosure for the sample to be transported, tested(e.g., cell count analysis, hemoglobin analysis, infectious diseasescreening, human leukocyte antigen (HLA) typing), and/or otherwiseprocessed. During these processing steps, and as described above withrespect to FIG. 7 , the vial 200 may be supported by the above-describedstorage container 700, and the tubes 300, 400 may be supported by thetube clip 500 and/or the spool 600.

The method of FIG. 8 continues with an opening step (e) with thecryovial device 100 in an opened configuration and a coupling step (f)with the cryovial device 100 in a coupled configuration. During theopening step (e), the inlet/outlet tube 300 is sliced along cut line 314below the seal 310, and the vent tube 400 is sliced along cut line 414below the seal 410 but still above the filter element 402 (FIG. 1 ). Inthis way, the inlet/outlet tube 300 becomes progressively shorter fromthe filling step (a), to the severing step (c), to the opening step (e).During the coupling step (f), the now-opened end of the inlet/outlettube 300 is coupled (e.g., welded) to a receiving tube R in a direct,closed manner. The opening step (e) and the coupling step (f) may beperformed substantially simultaneously in a closed environment to avoidleakage and/or contamination of the sample. For example, both theopening step (e) and the coupling step (f) may be performed using atubing welder that cuts and heats adjoining ends of the inlet/outlettube 300 and the receiving tube R, such as the CONNECT-FLEX® TPE TubingWelder available from Saint-Gobain. The opening step (e) and thecoupling step (f) of the inlet/outlet tube 300 may be performed abovethe height of the vent tube 400 to avoid interfering with the vent tube400. If necessary, the inlet/outlet tube 300 may be unraveled furtherfrom the spool 600 (FIG. 1 ) for added length and clearance above thevent tube 400.

The method of FIG. 8 concludes with a draining step (g) with thecryovial device 100 in a drained configuration. During the draining step(g), the sample is directed from the inlet/outlet opening 205 of thevial 200 (FIG. 1 ), through the inlet/outlet tube 300, and through thereceiving tube R, as indicated by arrow G. The draining step (g) may beperformed at atmospheric pressure, with air entering the vial 200 viathe reopened vent tube 400 and its corresponding filter element 402(FIG. 1 ). The withdrawn sample may be directed to its desired end use,such as laboratory testing or clinical administration. In this way, thesample travels through the same inlet/outlet tube 300 in oppositedirections during the draining step (g) and the above-described fillingstep (a). The drained cryovial device 100 may be discarded.

While this invention has been described as having exemplary designs, thepresent invention can be further modified within the spirit and scope ofthis disclosure. This application is therefore intended to cover anyvariations, uses, or adaptations of the invention using its generalprinciples. Further, this application is intended to cover suchdepartures from the present disclosure as come within known or customarypractice in the art to which this invention pertains and which fallwithin the limits of the appended claims.

What is claimed is:
 1. A cryovial device comprising: a vial configuredto hold a liquid sample; an inlet/outlet tube coupled to the vial andconstructed of a weldable polymer, the inlet/outlet tube having: afilled configuration in which the inlet/outlet tube is coupled to asource of the liquid sample; and a drained configuration in which theinlet/outlet tube is coupled to a receiving tube; and a vent tubecoupled to the vial.
 2. The cryovial device of claim 1, wherein theinlet/outlet tube is coupled to the vial at a first fitting and the venttube is coupled to the vial at a second fitting.
 3. The cryovial deviceof claim 2, further comprising a sealing element, the sealing elementincluding a first hole, the first hole receiving the inlet/outlet tubeand compressing the inlet/outlet tube around the first fitting, thesealing element including a second hole, the second hole receiving thevent tube and compressing the vent tube around the second fitting. 4.The cryovial device of claim 1, wherein the inlet/outlet tube isconstructed of a thermoplastic elastomer.
 5. The cryovial device ofclaim 1, wherein the inlet/outlet tube is welded to the receiving tubein the drained configuration.
 6. The cryovial device of claim 1, whereinthe inlet/outlet tube has a closed configuration between the filledconfiguration and the drained configuration.
 7. The cryovial device ofclaim 6, wherein the inlet/outlet tube is heat-sealed in the closedconfiguration.
 8. The cryovial device of claim 7, wherein inlet/outlettube is heat-sealed beyond an extension of the vent tube from the vial.9. The cryovial device of claim 6, wherein the vent tube is heat-sealedin the closed configuration.
 10. The cryovial device of claim 1, whereinthe inlet/outlet tube is shortened between the filled configuration andthe drained configuration.
 11. The cryovial device of claim 1, whereinthe liquid sample comprises a suspension of blood cells.
 12. A method ofusing a cryovial device including a vial and an inlet/outlet tube, themethod comprising the steps of: filling the vial with a liquid samplevia the inlet/outlet tube; closing the inlet/outlet tube after thefilling step; cryopreserving the sample in the vial after the closingstep; opening the inlet/outlet tube after the cryopreserving step;coupling the inlet/outlet tube to a receiving tube; and draining thesample from the vial into the receiving tube via the inlet/outlet tube.13. The method of claim 12, further comprising the step of unravelingthe inlet/outlet tube from a spool after the closing step.
 14. Themethod of claim 12, wherein the closing step comprises heat-sealing theinlet/outlet tube.
 15. The method of claim 12, further comprising thestep of severing the inlet/outlet tube after the closing step.
 16. Themethod of claim 12, wherein the coupling step comprises welding theinlet/outlet tube to the receiving tube.